Three skin specimens of the infected wound at t20h were processed for scanning electron microscopy. Skin preparation and infection was done as describe above. After the skin was incubated for 20 hours at 37°C in a moisture chamber, 8 mm biopsies were taken and fixed in a solution of 4% formaldehyde and 0.5% glutaraldehyde (in 50 mM HEPES) for 48 hours at room temperature. One of the sample duplicates was sliced right through the wound area with a scalpel in order to reveal the skin profile. All skin samples were afterwards washed in 50 mM HEPES, dehydrated in 30, 50, 70, 90, 95, 100, 100% ethanol, critical point dried, mounted on aluminum stubs, sputter coated with a 12 nm layer of gold-palladium and finally examined in the SEM (ZEISS 1530 Gemini, Carl Zeiss Microscopy GmbH, Germany) operating at 3 kV using the in-lens electron detector.
Gemini 1530
Manufactured by Zeiss
Sourced in Germany
The Gemini 1530 is a scanning electron microscope (SEM) manufactured by Zeiss. It is designed to provide high-resolution imaging of a wide range of materials and samples. The Gemini 1530 utilizes advanced electron optics to produce detailed, high-quality images at magnifications up to 1,000,000x.
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14 protocols using gemini 1530
1
Scanning Electron Microscopy of Infected Skin
2
SEM Analysis of Biofilm Samples
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The biofilm cultivation was performed as described above. A triplicate of 5 mm glass beads were washed by dipping in ddH2O and then fixed in a solution of 4% paraformaldehyde and 0.25% glutaraldehyde in 20 mM HEPES buffer for 48 h at 20°C. The biofilm of the second batch of glass bead triplicates was removed by sonication for 10 min as described before, then the bead was dip-washed in ddH2O and finally fixed in the before mentioned fixative. Subsequently, all samples were air-dried, mounted on a stub with adhesive carbon tape and Acheson silver (DAG1415M, Plano GmbH, Wetzlar, Germany), then sputter coated with a 12 nm layer of gold-palladium and examined in the SEM (ZEISS 1530 Gemini, Carl Zeiss Microscopy GmbH, Germany) operating at 3 kV using the in-lens electron detector.
3
SEM Imaging of Freeze-Dried Bacterial Beads
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Beads containing bacteria and biofilms were freeze-dried19 (link), fixed to a stub with sticky carbon tape, and then completely spray-sprayed with a 12-nm layer of gold. SEM (ZEISS 1530 Gemini, Carl Zeiss Microscopy GmbH, Germany) was used to examine and photograph the surface of the beads16 (link).
4
Microscopic Examination of A. baumannii Mutant
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The A. baumannii wild type strain and the ∆tol-pal mutant were grown on glass coverslips in 12 well plates in TSB medium at 37 °C for 24 h. Samples were fixed (1.0% paraformaldehyde, 2.5% glutaraldehyde in 50 mM HEPES) for 24 h; dehydrated in 30, 50, 70, 90, 95, 100, and 100% ethanol; critical point dried; mounted on aluminum stubs; sputter coated with a 12 nm layer of gold–palladium; and finally examined in the SEM (ZEISS 1530 Gemini, Carl Zeiss Microscopy GmbH, Germany) operating at 3 kV using the in-lens electron detector.
5
Comprehensive Materials Characterization Protocol
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The XRD patterns of all samples were
recorded with a PANalytical MPD diffractometer using Cu Kα radiation
(λ = 0.1541 nm), and the data were recorded from 10 to 70°
(2θ). Powder samples were examined by scanning electron microscopes
using a LEO (Zeiss) 1530 Gemini field-emission. TEM images were recorded
by a Philips/FEI Tecnai F20 S-TWIN TEM instrument operating at 200
kV. The UV–vis absorption spectra of the as-synthesized powdered
samples were measured using BaSO4 as a reference on a Shimadzu
UV-2450 spectrophotometer. Bandgap energies were calculated by the
analysis of the Tauc plots resulting from Kubelka–Munk transformation
of the absorption spectra. The absorption spectrum of the NGQD solution
was recorded by UV–vis–NIR spectroscopy (PerkinElmer
LAMBDA 750) at room temperature. FTIR spectroscopy was performed on
a PerkinElmer Spectrum 100 spectrometer using the typical potassium
bromide (KBr) pellet technique. The PL spectra were recorded at room
temperature by using a HORIBA LabRam HR spectrometer with the 325
nm line of the He–Cd laser as the excitation source.
recorded with a PANalytical MPD diffractometer using Cu Kα radiation
(λ = 0.1541 nm), and the data were recorded from 10 to 70°
(2θ). Powder samples were examined by scanning electron microscopes
using a LEO (Zeiss) 1530 Gemini field-emission. TEM images were recorded
by a Philips/FEI Tecnai F20 S-TWIN TEM instrument operating at 200
kV. The UV–vis absorption spectra of the as-synthesized powdered
samples were measured using BaSO4 as a reference on a Shimadzu
UV-2450 spectrophotometer. Bandgap energies were calculated by the
analysis of the Tauc plots resulting from Kubelka–Munk transformation
of the absorption spectra. The absorption spectrum of the NGQD solution
was recorded by UV–vis–NIR spectroscopy (PerkinElmer
LAMBDA 750) at room temperature. FTIR spectroscopy was performed on
a PerkinElmer Spectrum 100 spectrometer using the typical potassium
bromide (KBr) pellet technique. The PL spectra were recorded at room
temperature by using a HORIBA LabRam HR spectrometer with the 325
nm line of the He–Cd laser as the excitation source.
6
Quantifying Surface Charge of Nanocapsules
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The amount of surface charged groups was calculated from the results of the titration experiments performed on a Mütek particle charge detector (BTG, Herrsching, Germany) in combination with a Titrino Automatic Titrator (Metrohm AG, Herisau, Switzerland). The carboxylic groups were titrated against the positively charged polycation poly(diallyl dimethyl ammonium chloride) ( poly-DADMAC). The amine groups on the nanocapsules surface were titrated against the negatively charged polyelectrolyte poly(ethylene sulphonate) (PES-Na).
The titrations were performed on 10 mL of the nanocapsules dispersion with a solid content of 1 g L -1 . The amount of groups per gram of polymer was calculated from the consumed volume of the polyelectrolyte solution. Morphological studies were performed with scanning electron microscopy (SEM). The images were recorded by using a field emission microscope (LEO (Zeiss) 1530 Gemini, Oberkochen, Germany) operated at an accelerating voltage of 170 V. The samples were prepared by diluting the nanocapsule dispersion to about 0.01% solid content and by placing a droplet onto silica wafers and drying under ambient conditions.
The titrations were performed on 10 mL of the nanocapsules dispersion with a solid content of 1 g L -1 . The amount of groups per gram of polymer was calculated from the consumed volume of the polyelectrolyte solution. Morphological studies were performed with scanning electron microscopy (SEM). The images were recorded by using a field emission microscope (LEO (Zeiss) 1530 Gemini, Oberkochen, Germany) operated at an accelerating voltage of 170 V. The samples were prepared by diluting the nanocapsule dispersion to about 0.01% solid content and by placing a droplet onto silica wafers and drying under ambient conditions.
7
Backscattered Electron Imaging of Recovered Solids
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Backscattered electron (BSE) images of polished sections of the recovered solid phase were taken on a Zeiss Gemini 1530 field emission scanning electron microscope (FE-SEM) at Bayerisches Geoinstitut, University of Bayreuth. The accelerating voltage was 15–20 kV. The working distance was ∼14 mm and the aperture was 60 μm. An EDS was used to qualitatively identify mineral phases in the run products.
8
Nanocarrier Morphology by SEM
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The morphology of the nanocarriers was examined with a Gemini 1530 (Carl Zeiss AG, Oberkochem, Germany) scanning electron microscope (SEM) operating at 0.35 kV. The samples were prepared by casting a diluted and purified nanocarrier dispersion on silicon wafers.
9
Scanning Electron Microscopy of Biomass
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Selected biomass samples were studied by scanning electron microscopy (SEM). A small amount of freeze-dried (Flexi-dry, FTS systems, USA) biomass was fixed on conductive polycarbonate stickers with admixed graphite (G3347, Plano, Germany), and the samples were coated with a 3-nm-thick platinum layer in a sputter coater (MED 010, Bal-Tec, Liechtenstein). The electron microscope (Gemini1530, Zeiss, Germany) was operated at an acceleration voltage of 2 kV using a secondary electron detector [36 ].
10
Characterization of Fe, Mn, and Ca Minerals
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The appearance and formation of Fe, Mn and Ca minerals was investigated and characterised using a scanning electron microscope (type: Zeiss/Leo Gemini 1530, resolution 5 nm, at Ruhr-University Bochum) on 10 samples from the sediments of the drilled cores (1 sample from the Quaternary, 9 samples from the Tertiary aquifer). In addition, spectral analysis (wavelength-dispersive point measurements) was used to obtain chemical information of individual components.
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